Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c Mice
Treatment for toxoplasmosis is not completely successful because of their unwanted side effects, and new treatments are needed. Imiquimod has ability to moderate immune response and used to treat a wide variety of infections and tumors. The aim of the present study was to evaluate the effect of imiq...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2020-07-01
|
| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fcimb.2020.00387/full |
| _version_ | 1819205886019633152 |
|---|---|
| author | Leila Zaki Fatemeh Ghaffarifar Zohreh Sharifi John Horton Javid Sadraei |
| author_facet | Leila Zaki Fatemeh Ghaffarifar Zohreh Sharifi John Horton Javid Sadraei |
| author_sort | Leila Zaki |
| collection | DOAJ |
| description | Treatment for toxoplasmosis is not completely successful because of their unwanted side effects, and new treatments are needed. Imiquimod has ability to moderate immune response and used to treat a wide variety of infections and tumors. The aim of the present study was to evaluate the effect of imiquimod on the tachyzoites of T. gondii and infected macrophages in vitro and in BALB/c mice. The viability of T. gondii was assessed in the presence of various concentrations of imiquimod by direct counting after 6 and 24 h. The MTT assay was used to identify the viability of uninfected macrophages. The apoptotic effects were determined with flow cytometry on the tachyzoites and infected macrophages. For evaluation of parasite load in pre-treatment or post-treatment of macrophages Quantitative real time PCR (qPCR) was performed. For in vivo experiments, BALB/c mice received imiquimod before and after challenge with parasites. The mortality rate of mice, parasite numbers in spleen, and the INF-γ and IL-4 cytokine levels in spleen lymphocytes were evaluated. Imiquimod demonstrated anti-Toxoplasma effects by reducing the number of tachyzoites. The results of flow cytometry for drug-treated tachyzoites showed that apoptosis did not rise significantly relative to the control group (p < 0.05). Moreover, apoptosis was enhanced in infected macrophages as the concentration of imiquimod was reduced. The parasitic burden in imiquimod pretreated macrophages was significantly lower than those treated after infection (p < 0.01). A marked reduction was observed in survival rate, parasite load and INF-γ level in BALB/c mice that received imiquimod before parasitic challenge relative to those received drug after parasitic challenge (p < 0.01). Overall, imiquimod in the pretreated group had greater anti-Toxoplasma effects than imiquimod in posttreated group in vitro and in vivo. imiquimod may be considered as a candidate for use against Toxoplasmosis both therapeutically and prophylactically. |
| first_indexed | 2024-12-23T04:58:49Z |
| format | Article |
| id | doaj.art-e658ccb5c2e2444186724f2a5f4068ad |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-12-23T04:58:49Z |
| publishDate | 2020-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-e658ccb5c2e2444186724f2a5f4068ad2022-12-21T17:59:16ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882020-07-011010.3389/fcimb.2020.00387547240Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c MiceLeila Zaki0Fatemeh Ghaffarifar1Zohreh Sharifi2John Horton3Javid Sadraei4Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranDepartment of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranBlood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, IranTropical Projects, Hitchin, United KingdomDepartment of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranTreatment for toxoplasmosis is not completely successful because of their unwanted side effects, and new treatments are needed. Imiquimod has ability to moderate immune response and used to treat a wide variety of infections and tumors. The aim of the present study was to evaluate the effect of imiquimod on the tachyzoites of T. gondii and infected macrophages in vitro and in BALB/c mice. The viability of T. gondii was assessed in the presence of various concentrations of imiquimod by direct counting after 6 and 24 h. The MTT assay was used to identify the viability of uninfected macrophages. The apoptotic effects were determined with flow cytometry on the tachyzoites and infected macrophages. For evaluation of parasite load in pre-treatment or post-treatment of macrophages Quantitative real time PCR (qPCR) was performed. For in vivo experiments, BALB/c mice received imiquimod before and after challenge with parasites. The mortality rate of mice, parasite numbers in spleen, and the INF-γ and IL-4 cytokine levels in spleen lymphocytes were evaluated. Imiquimod demonstrated anti-Toxoplasma effects by reducing the number of tachyzoites. The results of flow cytometry for drug-treated tachyzoites showed that apoptosis did not rise significantly relative to the control group (p < 0.05). Moreover, apoptosis was enhanced in infected macrophages as the concentration of imiquimod was reduced. The parasitic burden in imiquimod pretreated macrophages was significantly lower than those treated after infection (p < 0.01). A marked reduction was observed in survival rate, parasite load and INF-γ level in BALB/c mice that received imiquimod before parasitic challenge relative to those received drug after parasitic challenge (p < 0.01). Overall, imiquimod in the pretreated group had greater anti-Toxoplasma effects than imiquimod in posttreated group in vitro and in vivo. imiquimod may be considered as a candidate for use against Toxoplasmosis both therapeutically and prophylactically.https://www.frontiersin.org/article/10.3389/fcimb.2020.00387/fullToxoplasma gondiiimiquimodmacrophagesin vivoin vitro |
| spellingShingle | Leila Zaki Fatemeh Ghaffarifar Zohreh Sharifi John Horton Javid Sadraei Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c Mice Frontiers in Cellular and Infection Microbiology Toxoplasma gondii imiquimod macrophages in vivo in vitro |
| title | Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c Mice |
| title_full | Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c Mice |
| title_fullStr | Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c Mice |
| title_full_unstemmed | Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c Mice |
| title_short | Effect of Imiquimod on Tachyzoites of Toxoplasma gondii and Infected Macrophages in vitro and in BALB/c Mice |
| title_sort | effect of imiquimod on tachyzoites of toxoplasma gondii and infected macrophages in vitro and in balb c mice |
| topic | Toxoplasma gondii imiquimod macrophages in vivo in vitro |
| url | https://www.frontiersin.org/article/10.3389/fcimb.2020.00387/full |
| work_keys_str_mv | AT leilazaki effectofimiquimodontachyzoitesoftoxoplasmagondiiandinfectedmacrophagesinvitroandinbalbcmice AT fatemehghaffarifar effectofimiquimodontachyzoitesoftoxoplasmagondiiandinfectedmacrophagesinvitroandinbalbcmice AT zohrehsharifi effectofimiquimodontachyzoitesoftoxoplasmagondiiandinfectedmacrophagesinvitroandinbalbcmice AT johnhorton effectofimiquimodontachyzoitesoftoxoplasmagondiiandinfectedmacrophagesinvitroandinbalbcmice AT javidsadraei effectofimiquimodontachyzoitesoftoxoplasmagondiiandinfectedmacrophagesinvitroandinbalbcmice |